Epstein Barr virus (EBV) is associated with 200,000 cases of cancer each year worldwide and is the cause of infectious mononucleosis. The virus is also associated with autoimmune diseases including multiple sclerosis. This year we developed a rapid assay to measure EBV antibodies in human sera. The assay involves incubation of sera with an EBV EBNA-luciferase fusion protein, adding paramagnetic protein A/G beads, collecting the beads with the immune complexes, adding luciferase substrate and measuring the light produced with a luminometer. The 45 second assay has 92% sensitivity and 100% specificity with virtually the same diagnostic performance as the FDA-approved commercial EBV VCA ELISA which takes several hours. The assay is also highly quantitative and requires 0.1 ul of sera to perform. This assay could be used as a point-of-care test for diagnosis of EBV infection or to identify seronegative volunteers for vaccine studies.